ABSTRACT: Biosynthesis is the only source of potato starch which is an important raw material for food processing, modified starch and biomass energy. However, it is not clear about the evolution of starch synthesis with tuber development in potato. The present study evaluated the differences of starch synthesis and gelatinization properties of potato tubers with different starch content. Relative to cultivars of medium and low starch content, cultivars of high starch content showed significantly higher SBEII gene expression, AGPase and SSS enzyme activity, and total starch content after middle stage of starch accumulation, and had smaller average starch granule size during whole process of tuber development, and had higher pasting temperature before late stages of tuber growth, and had lower pasting temperature after middle stage of starch accumulation. Path analysis showed that, after middle stage of starch accumulation, effects on starch gelatinization of cultivars with high, medium and low starch content represented starch synthesis enzyme activity > starch accumulation > starch granule distribution > starch synthesis enzyme gene expression, starch synthesis enzyme gene expression > starch synthesis enzyme activity > starch accumulation > starch granule distribution, starch synthesis enzyme gene expression > starch granule distribution > starch synthesis enzyme activity > starch accumulation, respectively. In the study, phases existed in the starch biosynthesis of potato tuber, and the starch quality and its formation process were different among varieties with different starch content. The findings might contribute to starch application and potato industries.